6x10ml Swing Rotor 

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V3-V4 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V3-V4 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V3-V4 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70400 (24 preps)	Cat. 70410, 70420, 70430, 70440 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V3-V4 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL

• For sizing and quantification of double strand DNA fragments.
• Composed of 5 bands from 10 kb to 48.5 kb.
• Premixed with 6X DNA loading buffer for direct gel loading.

• For sizing and quantification of double strand DNA fragments.
• Composed of 5 bands from 10 kb to 48.5 kb.
• Premixed with 6X DNA loading buffer for direct gel loading.

Product Description

Product Detail

Kit Storage and term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal nucleic acid Principle Summary

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39 ºC~42 ºC), reverse transcriptase uses specific primers and template RNA to synthesize cDNA strands, and binds the auxiliary protein and single strand With the help of the protein, the recombinase and the primer form a complex; perform a homology search and bind the target homology domain, at this time a D-loop region is formed at the homology position and strand exchange begins; accompanied by the recombinase from the complex Upon dissociation, the polymerase also binds to the 3′ end of the primer, initiating chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.

Isothermal nucleic acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.

Technical Parameters:

Parameters	Details
Product Name	RNA Isothermal Amplification Kit NFO
Manufacturer	Amp-future
Storage Temperature	-20°C
Kit Components	Enzymes, Buffers ,Reagents
Packaging	48 Tests/box
Detection Limit	500-1000copies/µL
Shipping	ICE
Test Time	5-20mins

Isothermal nucleic acid Applications

Suitable for RNA isothermal rapid amplification kit(NFO type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

RNA NFO kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.